This study focuses on the isolation, production and comparative analysis of α-amylase from strains of Bacillus and Aeromonas using three different habitats (cow dung dump waste site, fruit and vegetable waste dump site and rice field waste dump) site. Alpha amylase (α-1,4- glucan-glucanhydrolase, EC 3.2.1.1) is an extracellular enzyme that breaks down α-1,4 glycosidic linkages in starch and related substrates, producing oligosaccharides such as maltose, glucose, and alpha limit dextrin. Results showed that the α-amylase production by Bacillus subtilis strain was highest with 8.57 IU/ml/min and Aeromonas jandaei produced minimum amylase which is 4.80 IU/ml/min among all the six strains. Upon optimization of various parameters maximum production was observed at 24 hours to 48 hours, and the crude enzyme characterization revealed optimal activity at pH 7 and 37°C.